PDF(Pubmed)
Abstract：
Gene-modification therapies are at the forefront of HIV-1 cure strategies. Chimeric antigen receptor (CAR)-T cells pose a potential approach to target infected cells during antiretroviral therapy or following analytical treatment interruption (ATI). However, there are technical challenges in the quantification of HIV-1-infected and CAR-T cells in the setting of lentiviral CAR gene delivery and also in the identification of cells expressing target antigens. First, there is a lack of validated techniques to identify and characterize cells expressing the hypervariable HIV gp120 in both ART-suppressed and viremic individuals. Second, close sequence homology between lentiviral-based CAR-T gene modification vectors and conserved regions of HIV-1 creates quantification challenges of HIV-1 and lentiviral vector levels. Consideration needs to be taken into standardizing HIV-1 DNA/RNA assays in the setting of CAR-T cell and other lentiviral vector-based therapies to avoid these confounding interactions. Lastly, with the introduction of HIV-1 resistance genes in CAR-T cells, there is a need for assays with single-cell resolution to determine the competence of the gene inserts to prevent CAR-T cells from becoming infected in vivo. As novel therapies continue to arise in the HIV-1 cure field, resolving these challenges in CAR-T-cell therapy will be crucial.
摘要：
基因修饰疗法处于HIV-1治愈策略的最前沿。嵌合抗原受体(CAR)-T细胞在抗逆转录病毒疗法期间或在分析治疗中断(ATI)之后构成靶向感染细胞的潜在方法。然而,在慢病毒CAR基因递送的背景下,在HIV-1感染细胞和CAR-T细胞的定量以及在表达靶抗原的细胞的鉴定方面存在技术挑战。首先,缺乏有效的技术来鉴定和表征ART抑制和病毒血症个体中表达高变HIVgp120的细胞.第二,基于慢病毒的CAR-T基因修饰载体与HIV-1保守区之间的紧密序列同源性对HIV-1和慢病毒载体水平产生了定量挑战.需要考虑在CAR-T细胞和其他基于慢病毒载体的疗法中标准化HIV-1DNA/RNA测定，以避免这些混杂的相互作用。最后,随着在CAR-T细胞中引入HIV-1抗性基因，需要具有单细胞分辨率的测定法来确定基因插入物防止CAR-T细胞在体内被感染的能力。随着新疗法在HIV-1治愈领域的不断出现，解决CAR-T细胞治疗中的这些挑战将至关重要。
